Objective To investigate the relationship between cognitive function and sleep quality in patients with Alzheimer's disease(AD).Methods A case-control trial was conducted on 151 patients admitted to our department from May 2024 to December 2024.The patients with mild-to-moderate AD were assigned into the AD group(59 cases),and the cognitively normal patients in-to the control group(92 cases).All patients received neuropsychological assessment and sleep quality evaluation,including mini-mental state examination(MMSE),Montreal cognitive assess-ment(MoCA)and Pittsburgh sleep quality index(PSQI).The general clinical data and sleep qual-ity were compared between the two groups.Spearman correlation analysis was applied to analyze the relationship between cognitive function and sleep quality in the AD patients.Results The education level,MMSE score,MoCA score,instant story recall(ISR)score,delayed story recall(DSR)score,clock dawning test(CDT)score and Boston naming test score were significantly lower,while the activity of daily living(ADL)score was obviously higher in the AD group than the cognitively normal group(P＜0.05,P＜0.01).The AD patients had notably higher total score of PSQI and scores of sleep efficiency,nocturnal sleep disturbance and daytime dysfunction than the cognitively normal patients(P＜0.05,P＜0.01).In the AD patients,the total sleep time was positively correlated with the DSR score and the CDT score(r=0.300,P=0.021;r=0.308,P=0.018);hypnotic medication was negatively correlated with MMSE score,ISR score,and DSR score,and positively with ADL score(r=-0.320,P=0.013;r=-0.400,P=0.002;r=-0.331,P=0.010;r=0.355,P=0.006).Multiple linear regression analysis showed that benzodiazepines use and education level were independent influencing factors for cognitive function in the AD patients(P＜0.05,P＜0.01).Conclusion In AD patients,sleep quality is closely associated with cognitive function,as manifested by decreased sleep quality,reduced sleep efficiency,and worsened nocturnal sleep disturbances and daytime dysfunction.Administration of benzodiazepines may exacerbate cognitive impairment in AD patients.So,it is recommended that AD patients should be cautious about taking such drugs.

Objective To investigate the associations of immune cells,CD8+and CD4+T cells,and high-sensitivity C-reactive protein(hs-CRP)with cognitive function,and to explore the relation-ships among immunity,chronic inflammation,and Alzheimer's disease-related cognitive impair-ment.Methods A cross-sectional study was conducted on 101 patients with primary complaints of memory decline who visited the Alzheimer's Disease Clinic of Dongzhimen Hospital from June to December 2024.Mini-Mental State Examination(MMSE)and Delayed Story Recall Task(DSR)were performed to assess their cognitive function,and according to the results,they were divided into observation group(cognitively impaired,60 cases)and control group(cognitively normal,41 cases).Peripheral blood levels of CD8+T cells,CD4+T cells,and hs-CRP were compared between the two groups.Results The observation group exhibited significantly lower total scores and scores of different dimensions of MMSE and DSR scores,but notably higher activities of daily liv-ing scores than the control group(P＜0.05,P＜0.01).Serum hs-CRP level was obviously elevated in the observation group than the control group(P＜0.05).Binary logistic regression analysis revealed that CD8+T cells(OR=0.998,95％CI:0.996-1.000,P=0.038)and body mass index(OR=0.843,95％CI:0.719-0.990,P=0.037)were protective factors,while hs-CRP(OR=2.004,95％CI:1.215-3.306,P=0.006)was an independent risk factor for cognitive impairment.Spearman's rank correlation analysis showed a significant positive correlation between hs-CRP and CD4+T cells(P=0.011),but no significant association with CD8+T cells(P＞0.05).Conclusion Chronic inflammation and immune dysregulation synergistically contribute to cogni-tive decline.Hs-CRP may serve as a potential screening biomarker for cognitive impairment in pri-mary care settings.

ObjectiveTo explore the impact of exogenous chitosan on the growth and metabolism of Glycyrrhiza uralensis Fisch. (G. uralensis) and to improve the quality of cultivated G. uralensis for both medicine and food and aid in the increase in the content of effective components in G. uralensis.MethodsIn this study, whole G. uralensis plants were treated with exogenous chitosan, and comprehensive analyses of secondary metabolites and proteins were conducted using liquid chromatography with tandem mass spectrometry and isobaric tag for relative and absolute quantitation, respectively. Effects of chitosan induction on endogenous hormones of G. uralensis were analyzed using an enzyme-linked immunosorbent assay. Gene ontology function annotation and Kyoto Encyclopedia of Genes and Genomes pathway annotation were conducted to study the effect of chitosan induction on the proteome.ResultsChitosan induction significantly increased the levels of flavonoids in G. uralensis; however, the variation in triterpenoids was not substantial. Biological processes, including photosynthesis, secondary metabolism, and abiotic stress responses, were significantly enriched. Additionally, the photosynthetic pathway, photosynthesis-antenna protein pathway, and plant hormone signal transduction pathway were significantly enriched. In the flavonoid biosynthesis pathway, the upstream-related enzyme phenylalanine ammonia-lyase (PAL) and the downstream-related enzymes chalcone synthase (CHS), polyketide reductase (PKR), chalcone isomerase (CHI), and vestitone reductase (VR) were significantly upregulated.ConclusionsOur findings suggest that chitosan induction may promote the tricarboxylic acid (TCA) cycle, and the TCA cycle enhancement significantly upregulated PAL, CHS, PKR, CHI, and VR, the five key enzymes involved in flavonoid synthesis of G. uralensis, indicating that chitosan induction activated the entire metabolic pathway associated with flavonoids in G. uralensis. Our findings provide a reference for improving the quality of cultivated G. uralensis from the perspective of pharmacodynamic components.

The development of traditional Chinese medicine (TCM) diagnosis and treatment has undergone multiple paradigms, evolving from sporadic experiential practices to systematic approaches in syndrome differentiation and treatment and further integration of disease and syndrome frameworks. TCM is a vital component of the medical system, valued alongside Western medicine. Treatment based on syndrome differentiation embodies both personalized treatment and holistic approaches; however, the inconsistency and lack of stability in syndrome differentiation limit clinical efficacy. The existing integration of diseases and syndromes primarily relies on patchwork and embedded systems, where the full advantages of synergy between Chinese and Western medicine are not fully realized. Recently, driven by the development of diagnosis and treatment concepts and advances in analytical technology, Western medicine has been rapidly transforming from a traditional biological model to a precision medicine model. TCM faces a similar need to progress beyond traditional syndrome differentiation and disease-syndrome integration toward a more precise diagnosis and treatment paradigm. Unlike the micro-level precision trend of Western medicine, precision diagnosis and treatment in TCM is primarily reflected in data-driven applications that incorporate information at various levels, including precise syndrome differentiation, medication, disease management, and efficacy evaluation. The current priority is to accelerate the development of TCM precision diagnosis and treatment technology platforms and advance discipline construction in this area.

Objective To investigate the correlation between the expression of human leukocyte antigen class I(HLA-I)and programmed cell death ligand 1(PD-L1)with clinicopathological features and cellular immune infiltration.Methods A total of 150 patients with bladder cancer diagnosed and treated in Anhui Provincial Hospital from May 2020 to April 2023 were retrospectively selected as the study objects.The positive expression rates and positive scores of HLA-I and PD-L1 were compared between cancerous tissues and adjacent tissues.The positive scores of HLA-I and PD-L1 in cancer tissues of patients with different clinical characteristics were compared,and the correlation between HLA-I,PD-L1 and clinical characteristics of patients with bladder cancer was analyzed by Kendall's tau-b method.Logistic regression model was used to establish the combined model parameters of HLA-I and PD-L1 positive scores,and receiver operating characteristic(ROC)curve was drawn to analyze the HLA-I and PD-L1 positive scores and the area under the curve(AUC),sensitivity and specificity of the combined diagnosis of bladder cancer.Results The positive expression rate of HLA-I in cancer tissues was lower than that in paracancer tissues[38.67%(58/150)vs 81.33%(122/150)],while the positive expression rate of PD-L1 was higher than that in paracancer tissues[57.33%(86/150)vs 14.00%(21/150)],and the differences were statistically siginficant(χ2=56.889,61.377,all P<0.05).The HLA-I positive score of cancer tissues was lower than that of paracancer tissues[2.00(1.00,3.00)vs 3.00(3.00,5.00)],while the PD-L1 positive score was higher than that of paracancer tissues[3.00(2.00,5.00)vs 2.00(1.00,2.00)],and the fifferences were statistically significant(Z=-8.409,-6.346,all P<0.05).There was no significant difference in HLA-I and PD-L1 positive scores among different sex,age and tumor diameter(ZHLA-1=-1.834,-0.622,-0.543;ZPD-L1=0.811,0.812,0.919,all P＞0.05).The difference of HLA-I and PD-L1 positive scores among different pathological stages,lymph node metastasis,differentiation degree,CD4+,CD8+and CD68+were statistically significant(ZHLA-1=-7.034～3.814;ZPD-L1=-4.479～3.257,all P＜0.05).Kendall's tau-b correlation analysis showed that HLA-I was negatively correlated with pathological stage,lymph node metastasis,degree of differentiation,and positively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.528～-0.286,all P<0.05).PD-L1 was positively correlated with pathological stage,lymph node metastasis,degree of differentiation and negatively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.243～0.334,all P<0.05).ROC curve analysis showed that the positive scores of HLA-I and PD-L1 and the AUC values of the combined diagnosis of bladder cancer were 0.773,0.702 and 0.856,respectively.Sensitivity was 61.30%,57.30%and 82.00%.The specificity was 81.30%,86.00%and 73.30%.Conclusion The expression of HLA-I and PD-L1 is abnormal in patients with bladder cancer,and their expression is affected by the positive infiltration of immune cells.Observing the positive expression of HLA-I and PD-L1 is beneficial to provide guidance for clinical diagnosis and treatment.

OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.

Circular RNA(circRNA),as a kind of non-coding RNA,regulates a variety of biological functions.To explore the effect of circRNA on PEDV replication in the host porcine intestinal epi-thelial cells,this study screened and analyzed the differentially expressed circRNAs by bioinforma-tic software in African Green Monkey renal cells(Vero-E6 cells)infected by porcine epidemic di-arrhea virus(PEDV),the differentially expressed circRNA ssc_circ_PIK3C2A was identified and the secondary structure was analyzed.PCR was used to identify the ssc_circ_PIK3C2A circRNA structure,the model of PEDV-infected IPEC-J2 cells was constructed,the TCID50 test was used to validate the viral titer of PEDV.The expression of circ_PIK3C2A was detected by qRT-PCR in IPEC-J2 infected by PEDV.circ_PIK3C2A qRT-PCR was performed to detect the expression of N gene of PEDV when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.The results showed that ssc_circ_PIK3C2 A is a porcine circular RNA with a typical circular structure,the virus titer of PEDV reached 10-6/mL after PEDV infected IPEC-J2 cells for 48 h,the expression of circ_PIK3C2A increased extremely(P＜0.01)at 6 h after PEDV-infection,with the extension of infec-tion time,its expression gradually decreased,and the expression was the lowest at 24 h,but there was no time-dependent trend.The expression of PEDV N gene decreased significantly when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.In conclusion,when PEDV infects IPEC-J2 cells,the expression of porcine circ_PIK3C2A decreases,and replication of PEDV increases signifi-cantly in IPEC-J2 cells.our result provides a basis for further study of the mechanism of circular RNA on PEDV replication and its physiological activities in host cells in the future.

Transmissible gastroenteritis(TGE)is a highly contagious gastrointestinal disease of pigs caused by the transmissible gastroenteritis virus(TGEV).It results in severe diarrhea and high mortality in suckling piglets.As no specific treatment available,vaccination is considered to be one of the most cost-effective measures for preventing and controlling TGE.However,the immune protection provided by vaccines based on traditional TGEV strains is becoming inadequate due to the continuous emergence of strong and new strains of the virus,which poses a serious threat to the pig industry's health and development.Therefore,the development of new vaccines with im-proved efficiency and broad-spectrum coverage is urgently needed.This paper aims to summarize the pathogenic structural characteristics of TGEV,discuss the latest advancements in TGEV vac-cine research and development,and propose future strategies for the development of highly effec-tive TGEV vaccines.The findings of this paper can serve as a valuable reference for effectively pre-venting and controlling TGE in clinical practice.

Objective To investigate the correlation between the expression of human leukocyte antigen class I(HLA-I)and programmed cell death ligand 1(PD-L1)with clinicopathological features and cellular immune infiltration.Methods A total of 150 patients with bladder cancer diagnosed and treated in Anhui Provincial Hospital from May 2020 to April 2023 were retrospectively selected as the study objects.The positive expression rates and positive scores of HLA-I and PD-L1 were compared between cancerous tissues and adjacent tissues.The positive scores of HLA-I and PD-L1 in cancer tissues of patients with different clinical characteristics were compared,and the correlation between HLA-I,PD-L1 and clinical characteristics of patients with bladder cancer was analyzed by Kendall's tau-b method.Logistic regression model was used to establish the combined model parameters of HLA-I and PD-L1 positive scores,and receiver operating characteristic(ROC)curve was drawn to analyze the HLA-I and PD-L1 positive scores and the area under the curve(AUC),sensitivity and specificity of the combined diagnosis of bladder cancer.Results The positive expression rate of HLA-I in cancer tissues was lower than that in paracancer tissues[38.67%(58/150)vs 81.33%(122/150)],while the positive expression rate of PD-L1 was higher than that in paracancer tissues[57.33%(86/150)vs 14.00%(21/150)],and the differences were statistically siginficant(χ2=56.889,61.377,all P<0.05).The HLA-I positive score of cancer tissues was lower than that of paracancer tissues[2.00(1.00,3.00)vs 3.00(3.00,5.00)],while the PD-L1 positive score was higher than that of paracancer tissues[3.00(2.00,5.00)vs 2.00(1.00,2.00)],and the fifferences were statistically significant(Z=-8.409,-6.346,all P<0.05).There was no significant difference in HLA-I and PD-L1 positive scores among different sex,age and tumor diameter(ZHLA-1=-1.834,-0.622,-0.543;ZPD-L1=0.811,0.812,0.919,all P＞0.05).The difference of HLA-I and PD-L1 positive scores among different pathological stages,lymph node metastasis,differentiation degree,CD4+,CD8+and CD68+were statistically significant(ZHLA-1=-7.034～3.814;ZPD-L1=-4.479～3.257,all P＜0.05).Kendall's tau-b correlation analysis showed that HLA-I was negatively correlated with pathological stage,lymph node metastasis,degree of differentiation,and positively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.528～-0.286,all P<0.05).PD-L1 was positively correlated with pathological stage,lymph node metastasis,degree of differentiation and negatively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.243～0.334,all P<0.05).ROC curve analysis showed that the positive scores of HLA-I and PD-L1 and the AUC values of the combined diagnosis of bladder cancer were 0.773,0.702 and 0.856,respectively.Sensitivity was 61.30%,57.30%and 82.00%.The specificity was 81.30%,86.00%and 73.30%.Conclusion The expression of HLA-I and PD-L1 is abnormal in patients with bladder cancer,and their expression is affected by the positive infiltration of immune cells.Observing the positive expression of HLA-I and PD-L1 is beneficial to provide guidance for clinical diagnosis and treatment.